This project provides state-of-the-art research technologies for NIAID's intramural infectious diseases, allergy, and immunology research programs. The new technologies are developed and validated and then applied in support of NIAID research. Technologies developed outside the NIH are likewise tested, evaluated, validated and, if appropriate, incorporated into the technology portfolio of the NIAID intramural program. The technologies supported include flow cytometry, confocal microscopy, electron microscopy, DNA microarray, DNA sequencing, bacterial phenotyping and quantitative PCR. Many of these technologies are used in high containment laboratories critical to the Institute's infectious diseases and biodefense research agenda. In addition to technology development, the RTB provides advanced training in all aspects of the technologies in the Branch's portfolio. Sequencing The RTB develops applications using capillary DNA sequencing technology. Applications are developed in close collaboration with DIR investigators. All data is uploaded to a server, which tracks and manages all of the sequencing data. Phenotyping microarray The RTB develops applications using a phenotyping microarray technology. This technology has the capability to analyze >2,000 metabolic reactions for bacterial strains, clones, or tissue culture cells through the use of a 96-well plate format. Micro Arrays The RTB develops project-specific research applications on Affymetrix microarray platform including custom chip design, experimental design, sample processing and chip processing. In addition to developing new applications for microarray research, the RTB develops statistical analysis, data management, and data mining solutions for DIR research programs;focusing on interpreting data generated by highly parallel detection systems used in genomics. Human/Pathogen Genotyping Several technologies are used for human and pathogen genotyping depending on the scope of the genotyping project and they range from capillary-based re-sequencing of entire human genes for de novo SNP or In/Del discovery, to high throughput targeted SNP genotyping via allelic discrimination assays using Taqman quantitative PCR, to SNPlex multiplexed assays (3730XL sequencer), to Affymetrix SNP chip-based arrays and custom pathogen SNP (MIP technology) arrays. Flow Cytometry Project-specific research applications are developed for flow cytometry analysis and sorting in BSL-2 and BSL-3 environments. Electron Microscopy Project-specific research applications are developed in the areas of sample preparation and analysis ranging from basic structural studies to immuno-localization of selected antigens for a wide array of specimens. A variety of methods, protocols, and equipment are employed to accommodate different preparative and imaging needs. Recent technological advancements have primarily focused on introducing and optimizing sophisticated imaging and preparative technologies and techniques for improved retention and visualization of labile structures often lost during routine processing and improving structural preservation.